Lin X, Takahashi K, Campion S L, Liu Y, Gustavsen G G, Peña L A, Zamora P O
Medical Department, Brookhaven National Laboratory, Upton, NY, USA.
Int J Mol Med. 2006 May;17(5):833-9. doi: 10.3892/ijmm.17.5.833.
A multi-domain synthetic peptide, F2A4-K-NS, mimicked the action of recombinant human FGF-2 (rhFGF-2) in vitro and in an in vivo model of angiogenesis. Like rhFGF-2, F2A4-K-NS was quantitatively shown to bind to FGF receptors in a cell-free receptor binding assay using a chimeric FGFR1 (IIIc)/Fc as monitored by surface plasmon resonance (SPR), and also shown to bind to heparin using biotinylated low-molecular weight heparin in a similar SPR assay. In vitro, F2A4-K-NS triggered signal transduction as monitored by the stimulation of ERK1/2 phosphorylation in human umbilical cord endothelial cells. In cell based assays, it increased cell migration, cell proliferation, and gelatinase secretion; endpoints associated with FGF-2 stimulation. Furthermore, these in vitro effects were mediated with quantities of F2A4-K-NS that were similar to those of rhFGF-2. In vivo, F2A4-K-NS was angiogenic at doses of 40 and 400 ng/implant in a subcutaneous implant assay as determined by morphologic scoring, hemoglobin content, and histology. These results support the hypothesis that F2A4-K-NS is a mimetic of FGF-2 that can substitute for FGF-2 in vitro and in vivo. A synthetic mimetic of FGF-2, such as F2A4-K-NS, could be a useful tool in studying mechanisms of cell activation and potentially in various therapeutic applications.
一种多结构域合成肽F2A4-K-NS在体外和血管生成的体内模型中模拟了重组人FGF-2（rhFGF-2）的作用。与rhFGF-2一样，在使用嵌合FGFR1（IIIc）/Fc的无细胞受体结合试验中，通过表面等离子体共振（SPR）监测，定量显示F2A4-K-NS与FGF受体结合，并且在类似的SPR试验中，使用生物素化的低分子量肝素显示其与肝素结合。在体外，通过监测人脐静脉内皮细胞中ERK1/2磷酸化的刺激，F2A4-K-NS触发信号转导。在基于细胞的试验中，它增加了细胞迁移、细胞增殖和明胶酶分泌；这些终点与FGF-2刺激相关。此外，这些体外效应是由与rhFGF-2相似数量的F2A4-K-NS介导的。在体内，通过形态学评分、血红蛋白含量和组织学测定，在皮下植入试验中，40和400 ng/植入剂量的F2A4-K-NS具有血管生成作用。这些结果支持了F2A4-K-NS是FGF-2模拟物的假设，其可以在体外和体内替代FGF-2。FGF-2的合成模拟物，如F2A4-K-NS，可能是研究细胞激活机制以及潜在地用于各种治疗应用的有用工具。
